WO2004054977A1

WO2004054977A1 - Nicotinamide-based kinase inhibitors

Info

Publication number: WO2004054977A1
Application number: PCT/AU2003/001666
Authority: WO
Inventors: Christopher John Burns; Marcel Robert Kling
Original assignee: Cytopia Pty Ltd
Priority date: 2002-12-13
Filing date: 2003-12-15
Publication date: 2004-07-01
Also published as: EP1569907B1; CA2508171A1; EP1569907A4; EP1569907A1; CA2508171C

Abstract

A compound of the general formula (I) or pharmaceutically acceptable prodrugs, salts, hydrates, solvates, crystal forms or diastereomers thereof is described. A method of treating tyrosine kinase-associated disease states in a subject using a compound of formula (I) is also described.

Description

Nkotina-t-idc-based Kinase Inhibitors

FIELD OF THE INVENTION

The present invention involves compounds represented by Formula (I) herein below, phannaceutical compositions comprising Such compounds and methods of suppressing the growth of cancers and other proliferative diseases.

BACKGROUND OP THE INVENTION

Normal cellular proliferation is a well-controlled balance between the rate of cell cycle progression and programmed cell death (apoptosis). This balance is maintained by the appropriate transmission of extracellular signals by intracettular signal transduction circuitry- In tumours this equilibrium becomes disturbed by either unrestrained completion of the cell cycle, or loss of normal apoptotic cell death. In many cases this deregulation, comes about by the autonomous activation of the intracellular signal transduction circuitry that controls the cell cycle and apoptosis pathways. Central to the regulation of these pathways are members of the protein kinase family, and a. romising avenue to the generation of treatments for hyperproliferative diseases such as cancer, are compounds that target those kinases involved in this regulation.

Protein kinases are a family of enzymes that catalyse the phosphorylation of specific residues in proteins. In general protein kinases fall into several groups; those which preferentially phosphoiylate serine and/ or threonine residues, those which preferentially phosphorylate tyrosine residues and those which phosphorylate both tyrosine and Ser/ Thr residues. Protein kinases are therefore key elements in signal transduction pathways responsible for transducing extracellular signals, including the action of cytokines on their receptors, to the nuclei, triggering various biological events. The many roles of protein .kinases in normal cell physiology include cell cycle control and cell growth, differentiation, apoptosis, cell mobility and mitogenesis-

Inappropriately high protein kinase activity has been implicated in many diseases resulting from abnormal cellular function. This might arise either directly or indirectly, for example by failure of the proper control mechanisms for a kinase, related for example to mutation, over-expression or inappropriate activation of the enzyme; or by over- or underproduction of cytokines or growth factors also participating in the transduction of signals upstream or downstream of the kinase. In all of these instances, selective inhibition of tlie action of the kinase might be expected to have a beneficial effect. Diseases where aberrant kinase activity has been implicated include: diabetes; restenosis; atherosclerosis; fibrosis of the liver and kidney; ocular diseases; myelo- and lymphoproliferative disorders; cancer such as prostate cancer, colon cancer, breast cancer, head and neck cancer, leukemia and lymphoma; and, auto-immune diseases such as Atopic Dermatitis, Asthma, rheumatoid arthritis, Crohn's disease, psoriasis, Crouzon syndrome, achondroplasia, and thanatophoric dysplasia.

Protein kinases include, for example, but arc not limited to, members of the Protein Tyrosine Kinase family (PTKs), which in turn can be divided into the cytoplasmϊc PT s (CTKs) and the receptor PTKs (RTKs). The cytoplasmic PTKS include the SRC family, (including: BLK FGR; FYN; HCK; LCK; LYN; SRCYES and YRK); the BRK Family (inclu ing: BRK; FRK, SAD; and SEM); Hie CSK family (including: CSK and CTK); the BTK family, (including BTK; ITK; TEC; MKK2 and TXK), the Janus kinase family, (including: JAKI, JAK2, JAK3 and Tyk2), the FAK family (including, FAK and FYK2); the Fes family (including FES and FER), the 2AP70 family (including ZAP70 and SYK); the ACK family (including ACKl and ACK2); and the Abl family (including AB and ARC). The RTK family includes the ECF-Receptor family (including, EGFR, HER2, HER3 and HER4); the Insulin Receptor family (including INS-R and IGFl-R ); the FDGF-Receptor family (including PDGFRoc, PDGFRβ, CSF1R, KIT, FLK2 ); the VEGF-Receptor family (including; FLT1, F K1 and FLT4); the FGF-Receptor family (including PGFRt, FGFR2, FGFR3 and FGFR4 ); the CCK4 family (including CCK4); the MET family (including MET and RON); the TRK family (including TRKA, TRKB, and TRKC ); the AXL family (in ud g AXL, MER, and SKY); the TJE/TEK family (including TIE and ΪTE2/TEK); the EPH family (including EPHA1, EPHA2, EPHA3, EPHA4, EPHA5, EPHA6, EPHA7, EPHA8, EPHB1, EPHB2, EPHB3, EPHB , EPHB5, EPHB6); the RYK family (including RYK); the MCK family (including MCK and TYRO10); the ROS family (including ROS); the RET family (including RET); the LTK family (including LTK and ALK); the ROR family (including ROR1 and ROR2); The Musk family (including Musk); the IMR family including LMRl, LMR2 and LMR3); and the SuRTKl06 family (including SuRTKlOό). Similarly, the serine /threonine specific kinases (STKs) comprise a number of distinct subfamilies, including; the extracellular signal regulated kinases, (p42/ERK2 and p44/ERKI); c-Jun lMH2-terminal kinase (JNK); cAMP-responsive element-binding protein kinases (CREBK); cAMP-dependent kinase (CAPK); mitogen-activated protein kmase-activated protein kinase (MAPK and its relatives); stress-activated protein kinase p38/SAPK2; mitogen-and stress-activated kinase (MSK); protein kinases, PKA, PKB and PKC inter alia. Ad tionally, the genomes of a number of pathogenic organisms possess genes encoding protein kinases. For example, the malarial parasite Plasmodium falciparum and viruses such as HPV and Hepatitis viruses appear to bear kinase related genes.

In one embodiment, the method of the invention is used in the treatment of sarcomas, carcinomas and/ or leukemias. Exemplary disorders for which the subject method can be used alone or as part of a treatment regimen include: f-brosareoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogen sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocardnomas, cystadenocartinoma, medullary carcinoma, brαnchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocar noma, seminoma, embryonal cardnoma, Wilms' tumor, cervical cancer, testicular tumor, lung cardnoma, small cell lung cardnoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, ncuroblastoma, and retinoblastoiria.

In certain embodiments, the method of the invention is be used to treat disorders such as cardnomas forming from tissue of the breast, prostate, ki ney, bladder or colon.

In other embodiments, the method of the invention is used to treat hypeiplastic or neoplastic disorders arising in adipose tissue, such as adipose cell tumors, e,g,, lipomas, fibrolipomas, lipoblastomas, tipomatosis, hibemomas, hem-uigiomas and/ or liposarcomas. SUMMARY OF THE INVENTION

The present inventors have found that a group of compounds based upon a disubstitutcd pyridine scaffold are inhibitors of ihe growth and proliferation of cancer cells.

Accordingly, in a first aspect the present invention provides a compound of the general formula

I

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates, crystal forms or diastereomers thereof, wherein:

A is selected from O, S, NR1, where RI is selected from H, C_M alkyl;

B is aryl, hetaryl optionally substituted with 0-3 substituents independe tly chosen from halogen, .₄ alkyl, CF,, CN, aryl, hetaryl, OH, OCFa, OCwalk l, OC₂^alkyllMR2R3, Oaryl, Ohctaryl, COJR2, CONR2R3, NR2R3, C_M alkylNR2R3, NR4C_l4aIkyINR2R3, NR2COR3, OC(0)NR2R3, NR4CONR2R3, NR2S0₂R3; and R2,

R3 are each independently H, C^ alkyl, C_w alkyl heterocydyl, aryl, hetaryl, C_walkyl aryl, C_M alkyl hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR5; and R4 is selected from H, C_w alkyl; and R5 is selected from H, C_i alkyl;

Q is a bond, or C_]J( alkyl;

W is selected from H, C^alkyl, C_a.₆alkenyl; where C^alkyl or ^alkenyl may be optionally substituted with C_halky., OH, OC^alkyl, NRβC(0)R7, CONR6R7, OR6, NR6R7; and R6, and R7 are each independently H, C,.₄ alkyl, alkyl cycloalkyl, C1.4 alkyl heterocydyl, aryl, hetaryl, or may be joined to form an optionally substituted 3-8 mcmbered ring optionally containing an atom selected from O, S, NR8 and R8 is selected from H, C,.₄ alkyl;

Y is H, aryl or hetaryl optionally substituted with 0-3 substituents independently chosen from halogen, C^ alkyl, CF„ aryl, hetaryl, OH, OCF_v CN, C_._< alkynyl, OC_M alkyl, OC₂.₅alkylNR9R10, Oaryl, Ohetaryl, C0₂R9, CONR9R10, NR9R10, C_{l t} aϊkyINR9RlO, NRπC alkylNR9RlO, NR9COR10, NR11CONR9R10, NR95O₂R10; and R9, RIO are each independently H, C_w alkyl, C_{l l} alkyl heterocydyl, aryl, hetaryl, C_w alkyl aryl, C_w alkyl hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR12; and Rll is selected from H, C_w alkyl; and R12 is selected from H, C_M alkyl.

In a second aspect the present invention provides a composition comprising a carrier and at least one compound of the first aspect of the invention.

In a third aspect the present invention provides a method of treating a tyrosme kinase-associated disease state in a subject, the method comprising administering a therapeutically effective amount of at least one compound of the Hrst aspect of the invention or a therapeutically effective amount of a composition of the second aspect of the invention.

DETAILED DESCRIPTION OF TEJE INVENTION

In a first aspect the present invention provides a compound of the general formula

A is selected from O, S, NR1, where RI is selected from H, C,.₄ alkyl; B is aryl, hetaryl optionally substituted with 0-3 substituents independently chosen from halogen, C_w alkyl, CFs, CN, aryl, hetaryl, OH, OCF₃, OC^alkyl, OC₂.₅alkylNR2R3, Oaryl, Ohetaryl, CC₂R2, CONR2R3, NR2R3, C_M alkylNR2R3, NR4C,.₄alkylNR2R3, NR2COR3, OC(0)NR2R3, NR4CONR2R3, NR2SO₂R3; and R2, R3 are each independently FI, .₄ alkyl, C^ alkyl heterocydyl, aryl, hetaryl,

C_MaIkyI aryl, C_M alkyl hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR5; and R4 is selected from H, C alkyl; and R5 is selected from H, C_w alkyl;

Q is a bond, or C_w alkyl;

W is selected from H, C,.₄---kyl, -jalkenyl; where d^ l yl or C^alkenyi may be optionally substituted with C,.₄alkyl, OH, OC^alkyl, NR₆C(0)R7, C NR6R7, OR6, NR6R7; and R6, and R7 are each independently H, C,.₄ alkyl, C₁ alkyl cycloalkyl, C,.₄ alkyl heterocydyl, aryl, hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR8 and R8 is selected from H, C,.₄ alkyl;

Y is H, aryl or hetaryl optionally substituted with 0-3 substituents independently chosen from halogen, C,.₄ alkyl, CF₃, aryl, hetaryl, OH, OCF₃, CN, C_M alkynyl, O .₄ alkyl, O .₅atkyINR9R10, Oaryl, Ohetaryl, C0₂R9, CONR9R10, NR9R10, C_w alkylNR9RlO, NRll .₄alkylNR9R10, R9COR10, NRUCONR9R10, NR9SO₂R10; and R9, R10 arc each independently PI, C_M alkyl, .₄ alkyl heterocydyl, aryl, hetaryl, C_w alkyl aryl, C_1A alkyl hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR12; and Rll is selected from H, d-₄ alkyl; and R12 is selected from H, C_M alkyl.

In the above description it will be appredated that:

C_{1 £} alkyl means an unsubstituted or optionally substituted straight or branched alkyl chain

Aryl means unsubstituted or optionally substituted phenyl or naphthyl.

Hetaryl means an unsubstituted or optionally substituted 5- or 6-mcmbered heteroaromatic ring containing one or more heteroatoms selected from O, N, $. Cycloalkyl means a 3-8 membered saturated ring

Heterocydyl means a 3-8 membered saturated ring containing 1-3 heteroatoms selected from O, S, NR13, where R13 is H, C_tΛ alkyl, aryl, hetaryl.

Preferably, the compound is selected from the compounds of Table 1 and Table 2.

5 ϊn a further preferred embodiment the compound is selected from compounds of the general formula II.

II

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates, crystal forms or 10 diastereomers thereof, wherein:

RI is selected from H, C_H alkyl;

B is aryl, hetaryl optionally substituted with 0-3 substituents independently chosen from halogen, C alkyl, CFj, aryl, hetaryl, OH, OOP* OC,.₄alkyl, OC₂-₅alkylNR2R3, Oaryl, Ohetaryl CO_aR2, CONR2R3, NR2R3, C,.₄ alkylNR2R3, NR4C,-₊alkylNR2R3, 1.5 NR2COR3, NR4CONR2R3, NR2$0₂R3; and R2, R3 are each independently H, C_M alkyl, C_{l l} alkyl heterocydyl, aryl, hetaryl, C^al-kyl aryl, d.* alkyl hetaryl, or maybe joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR5; and R4 is selected from H, .₄ alkyl; and R5 is selected from H, C_..₄ alkyl;

20 Q is a bond, or Cι._Λ alkyl;

W is sdected from H, C_halky!, Q.₆alkenyl; where

may be optionally substituted with d.₄alkyl, OH, C,.₄alkyl, NR6R7; and R6, and R7 are each independently H, G^ alkyl, C,.₄ alkyl cycloalkyl, C_l4 alkyl h erocydyl, aryl, hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR8 and R8 is selected from H, C_w alkyl;

Y is H, aryl or hetaryl optionally substituted with 0-3 substituents independently chosen from halogen, C_H alkyl, CF<„ aryl, hetaryl, OH, OCF₃, OC^alky. , OC₂^alkylNR9R10, Oaryl, Ohctary-, CO₂R9, CONR9R10, NR9R10, C„ alkylNR9R10, NRllC,.₄alkylNR9R10, NR9COR10, NR11CONR9R10, NR9SO₂R10; and R9, RIO are each independently H, C_w alkyl, C_w alkyl heterocydyl, aryl, hetaryl, C^alkyl aryl, C_M alkyl hetaryl, or may be j ined to form an optionally substituted 3-8 membered ring optionally cont- tning an atom selected from O, S, NR12; and Rll is selected from H, C,.₄ alkyl; and R12 is selected from H, alkyl.

In the above description it will be appreciated that:

C,,₄ alkyl means an unsubstituted or optionally substituted straight or branched alkyl chain

Aryl means unsubstituted or optionally substituted phenyl or naphthyl-

Hetaryl means an unsubstituted or optionally substituted 5- or 6-membercd heteroaromatic ring cont-ώ-ting one or more heteroatoms selected from O, N, S.

Cycloalkyl means a 3-8 membered saturated ring

Heterocydyl means a 3-8 membered saturated ting containing 1-3 heteroatoms selected from O, S, NR13, where R13 is H, C_w alkyl, aryl, hetaryl.

The compounds of this invention include all conformational isomers (eg. ds and trans isomers). The compounds of the present invention may have asymmetric centers and therefore may exist in different enantiomeric and diastereomeric forms. This invention relates to the use of all optical isomers and stereoisomers of the compounds of the present invention, and mixtures thereof, and to all pharmaceutical compositions and methods of treatment that may employ or contain them. The compounds of formula I may also exist as tautomcrs. This invention relates to the use of all such tautomers and mixtures thereof.

This invention also encompasses pharmaceutical compositions containing prodrugs of compounds of the formula I. This invention also encompasses methods of treating or preventing disorders that can be treated or prevented by the inhibition of protein kinases comprising administering prodrugs of compounds of the formula I. Compounds of formula I having free amino, amido, hydroxy or carboxylic groups Can be converted into prodrugs. Prodrugs include compounds wherein an amino acid residue, or a polypeptide chain of two or more (eg, two, three or four) amino add residues which are covalently joined through peptide bonds to free amino, hydroxy and carboxylic acid groups of compounds of formula I. The amino add residues indude the 20 naturally occurring amino acids commonly designated by three letter symbols and also include, 4-hydroxyprolinc, hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvaline, beta-alanine, gamma-aininobutyric acid, dtrulHne, homocysteine, homoserine, omithine and methioine sulfone. Prodrugs also indude compounds wherein carbonates, carbamates, amides and alkyl esters which are covalently bonded to the above substituents of formula 1 through the carbonyl carbon prodrug sϊdediaϊn. Prodrugs also include phosphate derivatives of compounds of formula I (such as adds, salts of adds, or esters) joined through a phosphorus-oxygen bond to a free hydroxyl of compounds of formula I. Prodrugs also indude compounds wherem acyloxyalkyl or phosphonooxyalkyl moieties are covalently attached to compounds of formula I possessing a free hydroxyl group. Acyloxyalkyl or phosphonooxyalkyl moieties may also be covalen ly attached to compounds of formula I possessing a pyridyl ring through formation of a N-(acyloxyalkyi)- or N-(phosphonooxyalkyl)-pyridinium salt. This invention also encompasses pharmaceutical compositions containing prodrugs of compounds of the formula I.

In a third aspect the present invention provides a method of treating a tyrosine kinase-assodated disease state, the method comprising administering a therapeutically effective amount of at least one compound of the first aspect of the invention or a therapeutically effective amount of a composition of the second aspect of the invention.

In a preferred embodiment of the present invention the disease state is selected from the group consisting of Atopy, such as Allergic Asthma, Atopic Dermatitis (Eczema), and

Allergic Rhinitis; Cell Mediated Hypcrsensitivity, such as Allergic Contact Dermatitis and Hypersensitivity Pneumoni tis; Rheumatic Diseases, such as Systemic Lupus Erythematosus (SLE), Rlieumatoid Arthritis, Juvenile Arthritis, Sjδgren's Syndrome, Sderodcrma, Polymyositis, Ankylosing Spondylitis, Psoriatic Arthritis; Other autoimmune diseases such as Type I diabetes, autoimmune thyroid disorders, and Alzheimer's disease; Viral Diseases, such as Epstein Barr Virus (EBV), Hepatitis B, Hepatitis C, HIV, HTLV 1, Varicella-Zoster Virus (VZV), Human Papillo a Virus (HP V); Cancer, such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogem'c sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cardnoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell cardnoma, adenocardnoma, sweat gland carcinoma, sebaceous gland cardnoma, papillary cardnoma, papillary adenocardnomas, eystadenocardnoma, medullary carcinoma, bronchogenic cardnoma, renal cell carcinoma, hepatoma, bile duct cardnoma, choriocardnoma, semϊnoma, e bryonal cardnoma, Wilms' tumor, cervical cancer, testicular tumor, lung cardnoma, small cell lung cardnoma, bladder cardnoma, epithelial cardnoma, glioma, astrocytoma, medulloblastoma, cramopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, mei-ingioma, melanoma, neuroblastoma, and retinoblastoma, and car nomas foi-ming from tissue of the breast, prostate, kidney, bladder or colon, and neoplastic disorders arising in adipose tissue, such as adipose cell tumors, e.g., lipomas, fibrolipomas, lipoblastomas, Upomatosis, hibemomas, hemangiornas and/ or liposarcomas.

As used herein the term "tyrosine kinase-assodatcd disease state" refers to those disorders which result from aberrant tyrosine kinase activity and/or which are alleviated by inhibition of one or more of these enzymes.

The present invention provides pharmaceutical compositions comprising at least one of the compounds of the formula I or II capable of treating a kinase assόdated disorder in an amount effective therefore, and a pharmaceutically acceptable vehide or diluent. The compositions of the present invention may contain other therapeutic agents as described below, and may be formulated, for example, by employing conventional solid or liquid vehicles or diluents, as well as pharmaceutical additives of a type appropriate to the mode of desired administration (for example, exdpients, binders, preservatives, stabilizers, flavours, etc.) according to techniques such as those well known in the art of phannaceutical formulation. The compounds of the formula 1 or II may be administered by any suitable means, for example, orally, such as in the form of tablets, capsules, granules or powders; sublingually; buccally; p-u-enterally, such as by subcutaneous, intravenous, intramuscular, or mtradsternal injection or infusion techniques (e.g., as sterile injectable aqueous or non- aqueous solutions or suspensions); nasally such as by inhalation pray; topically, such as in the form of a cream or ointment; or redally such as in the form of suppositories; in dosage unit formulations containing non-toxic, pharmaceutically acceptable vehides or diluents. The compounds may, for example, be administered in a form suitable for immediate release or extended release. Immediate release or extended release may be achieved by the use of suitable pharmaceutical compositions comprising the present compounds, or, particularly in the case of extended release, by the use of devices such as subcutaneous implants or osmotic pumps.

In addition to primates, such as humans, a variety of other mammals can be treated according to the method of the present invention. For instance, mammals including, but not limited to, cows, sheep, goats, horses, dogs, cats, guinea pigs, rats or other bovine, ovine, equine, canine, feline, rodent or murine spedes can be treated. However, the method can also be practiced in other spedes, such as avian spedes (e.g., chickens).

Diseases and conditions assodated with inflammation and infection can be treated using the method of the present invention. In a preferred embodiment, the disease or condition is one in which the actions of εosinophils and / or lymphocytes arc to be inhibited or promoted, in order to modulate the inflammatory response.

The subjects treated in the above methods, in whom which cell growth inhibition is desired, are mammals, including, but not limited to, cows, sheep, goats, horses, dogs, cats, guinea pigs, rats or other bovine, ovine, equine, -anine, feline, ro ent or murine species, and preferably a human being, male or female-

The term "therapeutically effective amount" means the amount of the subject composition that will clidt the biological or medical response of a tissue, system, animal or human, that is being sought by the researcher, veterinarian, medical doctor or other clinidan-

The term "composition" as used herein is intended to encompass a product comprising the specified ingredients in the spedfied amounts, as well as any product which resul ts, directly or indirectly, from combination of the spedfied ingredients in the spedfied amounts. By "pharmaceutically acceptable" it is meant the carrier, diluent or exdpient must be compatible with the other ingredients of the formulation and not deleterious to the redpient thereof.

The terms "administration of" and or "administering a" compound should be understood to mean providing a compound of the invention to the individual in need of treatment.

The pharmaceutical compositions for the admi-n-stration of the compounds of this invention may conveniently be presented in dosage unit form and may be prepared by any of the methods well known in the art of pharmacy. AH methods include the step of bringing the active ingredient into association with the carrier, which constitutes one or more accessory ingredients. In general, the pharmaceutical compositions are prepared by uniformly and intimately bringing the active ingredient into assodation with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product into the desired formulation. In the pharmaceutical composition the active object compound is induded in an amount suffident to produce the desired effect upon the process or condition of diseases. As used herein, the term "composition" is intended to encompass a product comprising the spedfied ingredients in the spedfied amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the spedfied amounts.

The pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use maybe prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable exάpients, which are suitable for the manufacture of tablets. These exdpients may be for example, inert diluents, such as calάum carbonate, sodium carbonate, lactose, caldu phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn stardi, or aiginic add; binding agents, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic add or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal trad and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed- They may also be coated to form osmotic therapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed wi th an inert solid diluent, for example, calάum carbonate, caldum p osphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active materials in admixture with exdpients suitable for the manufacture of aqueous suspensions. Such exdpients are suspending agents, for example sodium carboxymcthylceilulose, methylcellulose, hydroxy-propylmethylcelluk.se, sodium alginate, polvvinyl-pyrrolidonc, gum tragacanth and gum. acacia; dispersing or wetting agents may be a naturaUy-occurring phosphatide, for example Iedthin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty adds and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty adds and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl, or n- propyl, p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such aβ liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic add.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional exdpients, for example sweetening, flavoring and coloring agents, may also be present.

The pharmaceu tJcal compositions of the invention may also be ϊn the form of oil-in-water emulsions. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin or mixtures of these. Suitable emulsifying agents may be naturally- occurring gums, for example gum acaria or gum tragacanth, naturally- occurring phosphatides, for example so bean, ledthin, and esters or partial esters derived from fatty adds and hexitol anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative and flavoring and coloring agents.

The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleagenous suspension. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injedable sol tion or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1,3-butane diol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed induding synthetic mono- or diglycerides. In addition, fatty adds such as oleic add find use in the preparation of injectables.

The compounds of the present invention may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating cxdpient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols. For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the compounds of the present invention are employed. (For purposes of this application, topical application shall indude mouthwashes and gargles.)

The compounds of the present invention can also be administered in the form of liposomes. As is known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by mono- or multilamellar hydrated liquid crystals that are dispersed in an aqueous medium. Any non-toxic, physiologically acceptable and metabolisable lipid capable of forming liposomes can be used. The present compositions in liposome form can contain, in addition to a compound of the present invention, stabilisers, preservatives, exdpients and the like. The preferred lipϊds are the phospholipids and phosphatidyl cholines, both natural and synthetic Methods to form liposomes are known in the art.

The pharmaceutical composition and method of the present invention may further comprise other therapeutically active compounds as noted herdn which are usually applied in the treatment of the above mentioned pathological conditions. Selection of the appropriate agents for use in combination therapy may be made by one of ordinary skill in the art, accordmg to conventional pharmaceutical prindples. The combination of therapeutic agents may ad svruergistically to effect the treatment or prevention of the various disorders described above. Using this approach, one may be able to achieve therapeutic efficacy with lower dosages of each agent thus reduάng the potential for adverse side effects.

Examples of other therapeutic agents indude the following:

cydosporins (e.g., cyclosporin A), CTLA4-Ig, antibodies such as ICAM-3, anti-IL-2 receptor (Anti-Tac), anti-CD45RB, anti-CD2, anti-CD3 (O T-3), anti-CD4, anti-CD80, anti-CD86, agents blocking the interaction between CD40 and gp39, such as antibodies spedfic for

CD40 and/or gp39 (i.e., CD154), fusion proteins constructed from CD40 and gp39 (CD401g and CD8gρ39), inhibitors, such as nudear translocation inhibitors, of NF-kappa B function, such as dooxyspergualin (DSG), cholesterol biosynthesis inhibitors such as HMG CoA reductase inhibitors (lovastatin and simvastatin), non-steroidal antiinflammatory drugs (NSAIDs) such as ibuprofen, aspirin, acetaminophen and cydooxygenase inhibitors such as rofecoxib, steroids such as prednisolone or dexamethasone, gold compounds, antiprolϊfera ive agents such as methotrexate, FK506 (tacrolimus, Prograf), m cophenolate mofetil, antineoplastic agents such as azathioprine, VP-16, etoposide, fludarabine, dspla m, doxorubid , adriamycin, amsacrine, camptothcdn, cytarabine, gemdtabine, viriblastine, vincristine, fluorodeoxyuridine, melphalan and eyclophosphamide, TNF-α inhibitors such as tenidap, anti-TNF antibodies or soluble TNF receptor, and rapamydn (sirolimus or Rapa unc) or derivatives thereof.

When other therapeutic agents are employed in combination with the compounds of the present invention they may be used for example in amounts as noted in the Physidan Desk Reference (FDR) or as otherwise determined by one of ordinary skill in the art.

The pharmaceutical composition and method of the present invention may further comprise other therapeutically active compounds as noted herein which are known inhibitors or substrates of drug efflux systems or drug detoxification and excretory systems. Such systems indude P-glycoprotein, multidrug resistance-assodated protein, lung resistance protein and glutathione $-transferase isoenzymes alpha, mu, pi, sig a, theta, zeta and kappa. Co-administration of drugs known to inhibit or reduce the activity of these systems may increase the efficacy of the compounds described in the present invention through increasing the amount of therapeutic agent in the cell. Using this approach, one may be able to achieve therapeutic efficacy with lower dosages, thus redudng the potential for adverse side effects. Examples of inhibitors or substrates for these systems indude; verapamil, probenecid, dipyridamole, ethacrynic add, indometha n, sulfasalazi e, buthioninc sulfoximine, cydosporin A and tamoxifcn.

In the treatment or prevention of conditions which require protein tyrosine kinase inhibition an appropriate dosage level will generally be about 0.01 to 500 mg per kg patient body weight per day which can be administered in single or multiple doses. Preferably, the dosage level will be about 0.1 to about 250 mg/kg per day; more preferably about 0.5 to about 100 mg/kg per day, A suitable dosage level may be about 0.01 to 250 mg/kg per day, about 0.O5 to 100 mg/kg per day, or about 0.1 to 50 mg/ g per day. Within this range the dosage ma be 0.05 to 0.5, 0.3 to 5 or 5 to 50 mg/kg per day. For. oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0, 10,0, 15.0. 20.0, 25.0, 50.0, 5.0,

100.0, 150.0, 200.0, 250.0, 300.0, 00.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. The compounds may be administered on a regimen of ϊ to 4 times per day, preferably once or twice per day.

It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors induding the activity of the speάfic compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

Throughout this spedfication the word "comprise", or variations such as "comprises" or "comprismg", will be understood to imply the mdusion of a stated clement, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

All publications mentioned in this pedfication are herein incorporated by reference.

Any discussion of documents, acts, materials, devices, artides or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed in Australia before the priority date of each claim of this application.

In order that the nature of the present invention may be more clearly understood preferred forms thereof will now be described by reference to the following non-limiting Examples.

EXAMPLES

MATERIALS AND METHODS:

Compound Syntiiesis

Compounds are generally prepared in a 2-step process starting from a protected 5- bromonicotinic acid. The first step of the synthesis typically involves a palladium mediated cross-coupling of the proterted 5-bromonicotinic add wi th a suitably funciionalised coupling partner. Typical coupling partners arc boronic acids (Suzuki coupling: see for example Miyaura, N. and Suzuki, Chem Rev.1995, -252457) or organostannanes (Stille coupling: see for example StiUe, J.K., Angew. Chem., Int. Ed. EngL, 19S6, 25, 508) (Scheme 1).

Scheme 1

The Suzuki coupling is the preferred coupling method and is typically performed in a solvent such as DME, THF, DMF, ethanol, propanol, toluene, or 1,4-dioxane in the presence of a base such as potassium carbonate, sodium carbonate, li hium hydroxide, caesium carbonate, sodium hydroxide, potassium fluoride or potassium phosphate. The reaction may be carried out at elevated temperatures and the palladium catalyst employed ma be selected from Pά P Pd(OAc);>, [PdCl₂(dppf)], Pd_(dba)₃/P(t-Bu)₃, palladium on carbon.

Methods to protect 5-bromonicotinic add are known to those skilled in the art and may include the formation of a 2-(trimethylsilyl)ethyl ester, 2-(t-±methyIsilyl)ethoxymethyl ester, tetrahydrofuranyl ester or £butyl ester. The £buiyl ester is tlie preferred protecting group.

The f"Butyl 5-bromo.nicotinate employed in the first s ep can be readily prepared from commercial 5-bromonicotinic add using conventional methods well known to those skilled in the art These methods indude the coupling of 5-bromonicotinic acid with t-butanol using coupling reagents such as dicydohexylcarbodiimide or l-(3-dimethylaminopropyl)- 3-ethylcarbodiimide in a solvent such as dichloromethane, or treatment of 5- bromonlcotinic add with di-f-butyldicarbonate in the presence of a base such as triethyla ine in a solvent such as tetrahydrofuran.

The second step of the synthesis is amide, ester or thioester formation by coupling the 5- arylnicotinic acid with a primary or secondary amine, an alcohol or phenol, or an alkyl or aryl mercaptan (Scheme 2).

Scheme 2

Initially the proterted ester forms of the 5-aryInicotinate derivatives prepared as in Scheme 1 are cleaved to the corresponding adds using methods well known to those skilled in the art.

The 5-arylnicotinic add derivatives are typically coupled with amines, alcohols, phenols, thiols or thiophenols using coupling reagents such as dicyclohexylca bodiimide, l-(3- dimethyIaminoproρy1)-3-ethylcarbodiimide, dϋsopropylcarbodiimide or carbonyldiimidazole ϊn solvents such as dichloromethane and tetrahydrofuran.

Alternatively, the deproterted 5-aryInicotinic add derivatives prepared as in Scheme 1 can be converted to the respective add chloride derivatives using thionyl chloride or oxalyl chloride, or to the mixed anhydride spedes using, for example, f-butyl chloroformate, using procedures well known to those skilled in the art. The add chloride or mixed anhydride derivatives can then be reacted with the desired amine, alcohol, phenol, thiol or thiophenol in the presence of a base such as triethylamine, diisopropylethylamine or solid phase equivalent in a solvent sudi as dichloromethane, tetrahydrofuran, dioxane or ethyl acetate at ambient or elevated temperatures, to generate the desired 5-aryl mcotinic add derivatives.

As a further alternative, the deproterted 5--uyInϊcotinic add derivatives prepared as in Scheme 1 can be converted to the corresponding active ester intermediates, such as the succini idyl, pentafluorophenyl or / nitrophenyl esters. This can be achieved by coupling the 5-aryInicotinic add derivatives with ΛAhydroxysucdnimide, pentafluorophenol orp- nitrophenol using coupling reagents such as dicyclohe-cylcarbodii i e, l-(3- dimethyl-uι-inopropyl)-3-ethylcarbodiimide, diisopropylearbodiimide or carbonyldiimidazole in solvents such as dichloromethane and tetrahydrofuran. Active acyl intermediates can also be formed directly by reaction of the 5-aryInicotinic add derivatives with reagents such as diphenylphosphoryl azide, pentafluorophenyl acetate. pentafluorophenyl diphenylphosphinate or cyanuric chloride using methods well known to those skilled in the art.

The products formed from this reaction sequence may be further derivatised using techniques well known to those skilled in the art,

Example 1

Terf_'Buiyl 5-b∑omomεaiinate

To a mixture of 5-bromonicotinic add (0.30g, 1.49mmol) and dϊ-terf-butyldicarbonate (0.45g, 2.06mmol) in THF (15mL) was added triethylamine (0.25mL, 1.79mmol) followed by 4-(pyrrolidino)pyridinc (30mg, 0.20mmol). Ihe resultant solution was stirred at ambient tomperature for 48 hr. The volatiles were then removed under vacuum and the residue was purified by column chromatography on silica (gradient, CH₂C1₂ to 3% MeOH/CH₂Cl₂) to provide the product as a white solid (0.35g, 91%)

'H-n.m.r. (CDC1₃) 51.60 (s, 9H t utyl), 8.35 (m, IH, Ar), 8.79 (d,J25 Hz, IH, Ar), 9.06 (d,/ 1.4 Hz, IH, Ar). Example 2.

Ter Btttyl5-(3,4- eihylenβctiaxypheflyl)tticotinate

In a flask was placed tert-butyl 5-bromonicotinate (l.OOg, 3.87mmoI), 10% w/w palladium on carbon (210mg, ~0.2mmol Pd), potassium carbonate (1.10g, 7.96mmol), 3,4- methylcnedioxy-phenyl boronic acid (0.96g, 5.79mmol), dimethylformamide (50mL) and water (250L). The flask was purged with nifrogen and then heated, with stirring, to 90°C for 15hr. The reaction mixture was allowed to cool, diluted with water (350mL) and extracted with CH₂C1₂ (4X). The combined extracts was washed with water, dried (MgSO and concentrated in vacuo. The crude residue was purified by flash chromatography on silica (gradient, CH,C1₂ to 50%ether/ CH₂C1₂) to provide an off-white solid (l.02g, 58%).

Η-n,m,r. (CDCL,) δl.63 (s, 9H, -butyl), 6.04 (s, 2H, CHJ, 6.90 (m, IH, Ar), 7.08 (m, 2H, Ar), 8.34 (m, IH, Ar), 8.89 (d, /1.9 Hz_r IH, Ar), 9.09 (d, /2.0 Hz, IH, Ar).

Example 3 5-(3, -methylenedioxyphenyl)mcotinoyI chloride

A solution of tert-bxAyl 5-(3,4-methylenedioxyphenyI)nicotinate (0.80g, 2.67mmol) in trifluoroacetic add (10mL) was stirred at room temperature for 2hr. The trifluoroacetic add was removed In vacuo and the residue treated twice with toluene followed by removal under vacuum (to remove residual TFA). The yellow/green residue was then treated with thionyl chloride (lO L) and dimethylformamide (ΪOOL) and the mixture was heated to reflux for 15hr. The reaction mixture was allowed to cool to ambient temperature, then the Volatiles were removed on a water aspirator. The residue was treated twice with toluene followed by removal in racαo to leave a yellow solid. To the crude add chloride was added 1,4-dioxane (16mL) to make a 0.167M suspension, which was used without further treatment in the next step.

Example 4

Formation of nicotinamides in 96-well format.

To each well of a 96-well deep well plate was added Amberlyst A-21 resin (70mg, 0.33mmol). To each well was then added a 0.19M 1,4-dioxane solution of amine (0.30mL, 57mol) followed by a 0.167M suspension of 5-atylnicotinoyl chloride (0-48mL, SOmol). The plate was sealed with a. webseal mat and the plate was sonicated for lhr in a sonicator bath. Amberlite IRA-67 (30mg, 0.17mmol) was added to each well, the plate was re-sealed and sonication continued for a further 3Qmin. The contents of the 96-well plate was transferred via pipettor to a 96-well filter plate and filtered into a second 96-well deep well plate. 1,4^ Dioxane (0.40mL) was placed in each well of the original %-wdl plate (to rinse). This was aspirated with the pipettor and then transferred to the filter plate and filtered into the second 96-well plate. The second 96-well plate was stripped of all volatiles on a Christ rotary vacuum concentrator, and then the residues in each well were re-dissolved in CH3CI2 (0.50mL). These CH₂CI₂ solutions were transferred to a second filter plate loaded with silica (200mg/ well) and filtered into a third 96-well deep well plate. A 10% methanol/ CH₂CI₂ solution (0.50mL) was added to each well of the second filter plate and filtered into the third deep well plate. The contents of the third plate was analysed by LC- MS and then concentrated under vacuum using the Christ rotary vacuum concentrator.

Further examples of 5-arylnicotinamides arc shown, in Table 1 along with their experimental m / z values. Table 1

Table 1 (cont)

Table 1 (cont)

Table 1 (cont.)

Table 1 (cont.)

SCREENING

JA Tyrosine Kinase Domain Production

fAK kinase domains were produced in the following manner:

JAKl

The kinase domain of human JAKl was amplified from U937mRNA using the polymerase chain reaction with the following primers;

XHOI-J1 5'-CCG CTC GAG ACT GAA GTG GAC CCC ACA CAT-3'

Jl- FNI 5'-CGG GGT ACC TTA TTT TAA AAG TGC TTC AAA-3'

JAKl PCR products were cloned into the pFastBac HTb expression vector (Gibco) via the Xho I and Kpn I sites. The JAKl plasmid was then transformed into competent DHlOBac cells (Gibco), and the recombinant baculovirus produced prepared for transfection into Sf9 insect cells.

JAK2

The kinase domain of humanJAK2 was amplified from U937πιRNA using the polymerase chain reaction with the following primers:

SALI-jk2 5'-ACG CGT CGA CGG TGC CTT TGA AGA CCG GGA T-3'

jk2-NOTT 5'-ATA GI TAG CGG CCG CTC AGA ATG AAG GTC ATT T-3'

JAK2 PCR prod cts were cloned into the pFastBac HTc expression vector (Gibco) via the Sal I and Not I sites. The JAK2 plasmid was then transformed into competent DHlOBac cells (Gibco), and the recombinant baculovirus produced prepared for transfection into Sf9 insect cells.

JAK3

The kinase domain of humanj AK3 was amplified from U937mRNA using the polymerase chain reaction with the following primers: XHOI-J3 5'-CCG CTC GAG TAT GCC TGC CAA GAC CCC ACG-3'

J3-KPNI 5'-CGG GGT ACC CTA TGA AAA GGA CAG GGA GTG-3'

JAK3 PCR products were cloned into the pFastBac HTb expression vector (Gibco) via the Xho I and Kpn I sites. The JAK3 plasmid was then transformed into competent DHlOBac cells (Gibco), and the recombma t baculovirus produced prepared for transfection into Sf insect cells.

TYK2

The kinase domain of humanTYK2 was amplified from A549 mRNA using the polymerase chain reaction with the following primers:

HT2EK 5'-GGA GCA CTC GAG ATG GTA GCA CACAAC CAG GTG-3'

ITY2.2R 5'-GGA GCA GGA ATT CCG GCG CTG CCG GTC AAA TCT GG-3'

TYK2 PCR products were cloned into pBIueBacHts2A (Invitrogen) via the ECQRI site. The recombinant TYK2 baculovirus produced was prepared for transfected into Sf9 insect cells.

Large Scale Production Of Kinase Domains

Baculovirus preparations from each of the JAK family members were infected into five litres of High Five cells (Invitrogen) grown in High Five serum free medium (Invitrogen) to a cell density of approximately 1-2 10* cells /ml. Cells are infected with virus at a MOI of 0.8-3.0 Cells were harvested and lysed. JAK kinase domains were purified by affinity chromatography on a Probond (Invitrogen) nickel chela te affinity column.

Assay Protocols

Kinase assays were performed either in a 96 well capture-based ELΪSA assay or in 384 well Optiplates (Packard) using an Alphascreen Protein Tyrosine Kinase kit. In either casse using approximately 1.5 μg of affinity purified PTK domain in the presence of 50rnM HEPES, pH 7.5, lOmM MgCI ISOmM NaCI and lOμM-lmM ATP. The biotinylated substrate biotin-EGPWLEEEEEAYG MDF-NH₂ (final concentration 5μM) was used as substrate. In the ELBA assay tyrosine phosphorylation was quantitated following transfer to an avidin coated E1ISA plate using peroxidase-linked anti-phospho-tyrosine antibody PY20, In the Alphascreen assay, Alphascreen phosphotyrosine acceptor beads followed by streptavidin donor beads were added under subdued light. The ELISA plates were read on a BMG Fluorostar, the Alphascreen plates were read on a Packard Fusion Alpha. Inhibitors were added to the assays fifteen minutes prior to the addition of ATP. Inhibitors were added in aqueous DMSO, with DMSO concentrations never exceeding 1%.

Results

The activity of a range of compounds is shown in Table 2. Compounds that exhibited a capacity to inhibit 50% or greater of enzyme activity at a concentration of 10 μM (measured under standard conditions, see Methods), are designated as "+". Compounds not tested are designated "NT"; while compounds that did not inhibit enzyme activity by 50% at 10 μM are designated "-".

3D

Table 2

Table 2 (cont)

Table 2 (cont)

Table 2 (cont)

Table 2 (cont.)

Table 2 (cont)

Table 1 (cont)

Table 2 (con*.)

Table 2 (cont)

Table 2 (cont)

Table 2 (cont)

Table 2 (cont.)

Table 2 (cont)

Table 2 (cont)

Table 2 (cont-)

Table 2 (cont)

Table 2 (cont)

Table 2 (cont)

Table 2 (cont)

Table 2 (cont)

Table 2 (cont)

Table 2 (cont)

S3

Table 2 (cont)

M

Table 2 (cont)

Table 2 (cont)

Table 2 (cont)

Table 2 (cont)

It will be appreciated by persons skilled in the art that numerous variations and/ or modifications may be made to the invention as shown in the specific embodiments without departing from the spirit or scope of the invention as broadly described. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.

Claims

CLAIMS:-

1 A compound of the general formula

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates, crystal fonns or diastereomers thereof, wherein:

A Is selected from O, S, NR1, where RI is selected from H, C_H alkyl;

B is aryl, hetaryl optionally substituted with 0-3 substituents independently chosen from halogen, C_M alkyl, CEj, CN, aryl, hetaryl, OH, OCFa, OC_Malkyt, OC₂^alkyINR2R3, Oaryl, Ohetaryl, CO^, CONK2R3, NR2R3, C_w alkylNR2R3,

and R2, R3 are each independently H, C_i alkyl, C^ alkyl heterocydyl, aryl, hetaryl, Cnalkyl aryl, C_M alkyl hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally cont--ining an atom selected from O, S, NR5; an R4 ts selected from H, C_w alkyl; and R5 is selected from H, C_{l l} alkyl;

Q is a bond, or -j alkyl;

W is selected from H, C_halky., C₂-₆alkenyl; where ^a-kyl or C^alkenyl may be optionally substituted ith C^alkyl, OH, OC alkyl, N ^O)--^, CONR6R7, OR6, NR6R7; and R6, and R7 are each independently H, C_w alkyl, C_w alkyl cycloalkyl, _w alkyl heterocydyl, aryl, hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NRS and R8 is selected from H, C₁ alkyl;

Y is I , aryl or hetaryl optionally substituted with 0-3 substituents independently chosen from halogen, C_w alkyl, CR„ aryl, hetaryl, OH, OCF₃, CN, C^ alkynyl, OC,.₄ alkyl, OC₂.₅aIkylNR9R10, Oaryl, Ohetaryl, CO₂R9, CONR9R10, NR9R10, .₄ alkylNR9R10, NRllC_1→alkyINR9R10, NR9COR10, NR11CONR9R10, NR S ₃RIO; and R9, RIO are each independently H, C_w alkyl, C_w alkyl heterocydyl, aryl, hetaryl, C,.₄ alkyl aryl, C_M alkyl hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR12; and Rll is selected from H, C^ alkyl; and R12 Is selected from H, C_M alkyl.

2. A compound according to claim 1 of tlie general formula II:

II

Rl is selected from H, C_w aϊkyl;

B is aiyl, hetaryl optionally substituted with 0-3 substituents independently chosen from halogen, C_M alkyl, CF₃ aryl, hetaryl, OH, OCF₃, OC^alkyl, ^I LNl-SRS, Oaiyl, Ohetaryl, C0₂R2, CONR2R3, NR2R3, -, alkylNR2R3, NR4C_walkylNR2R3, NR2COR3, NR4CONR2R3, NR2SO₂R3; and R2, R3 are each independently H, C_M alkyi, d^ alkyl heterocydyl, aryl, hetaryl, C_halky! aryl, C_t.₄ alkyl hetaryl, or maybe joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR5; and R4 is selected from H, C_{j l} alkyl; and R5 is selected from H, C_M alkyl;

Q is a bond, or C,.₄ alkyl;

W is selected from H, C_MaIkyI, -^alkenyl; where C_walkyl or C_?.._βalkenyl may be optionally substituted with C_MalkyI, OH, OC,.₄alkyI, NR6R7; and R6, and R7 are each independently H, C™ alkyl, C,.₄ alkyl cycloalkyl, -₄ alkyl heterocydyl, aryl, hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR8 and R8 is selected from H, .₄ alkyl; Y is H, aryl or hetaryl optionally substituted with 0-3 substituents independently chosen from halogen,, C_u alkyl, CF^ aryl, hetaryl, OH, OCFjj, OC^alkyl, OC₂^alkylNR9R10, Oaryl, Ohetaryl, C0₂R9, CONR9R10, NR9R10, C„ aIkyESrR9RlU, NRllC,.₄alkylNR9RlO, NRθCORlO, NR11CONR9R10, NR95O₂R10; and R9, RIO are each independently H, -j alkyl, C_w alkyl heterocydyl, aryl, hetaryl, C_MalkyI aryl, C_..₄ alkyl hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR12; and Rll is selected from H, C_w alkyl; and R12 is selected from W, C,.₄ alkyl.

A compound according to daim 1 wherein the compound is selected from the group consisting of:

C22H22N204 C23H230IN2O4

C20H18N2O2 C--2H20CIFN2O4

C19H15FN202 C22H20CIFN2O4

C19H16N202 C22H21CIN204

C--0H19N3O2 C20H16N2O3

C23H24N204 C16H12N20S

C1SH18N203 013H14N2OS

C--2H22N202 C18H16N20S

C21H19FN202 C20H19N3O2S

C21H19FN202 C1SH15N20S

C21H20M2O2 C17H13FN20S

C17H13FN20S C21H--0N2O2

C21H20N2O2

C20H20N2OS C22H22N203

C17H14N20S C21H20N--Q2

C1SH15FN20S C--0H17FN2O2

C19H17N303 C20H17FN2O2

C19H17N302 G20H15N2O2

C21H20N2O3

C22H2--N204 C20H17FN2O3

C18H16N203 C20H17FN2O3

C21H20N2O3 C20H18N2O3

4. A composition comprising a carrier and at least One compound of any one of daims 1 to 3.

5. A composition comprising a carrier and at least one compound of any one of claims l o .

6. A method of treating a tyrosine kinase-associated disease state in a subject_, the method comprising administering a therapeutically effective amount of at least one compound of any one of claims 1 to 4 or a therapeutically effective amount of a composition of claim 5.

7. A method according to daim 6 wherein the disease state is selected from the group consisting of Atopy, such as Allergic Asthma, Atopic Dermatitis (Eczema), and

Allergic Rhinitis; Cell Mediated Hypersensitiviiy, such as Allergic Contact Dermatitis and Hypersensitiviiy Pneumonitis; Rheumatic Diseases, such as Systemic Lupus Erythcmatosus (SLE), Rheumatoid Arthritis, Juvenile Arthritis, Sjogren's Syndrome, Scleroderma, Rolymyosiϋs, Ankylosing Spondylitis, Psoriatic Arthritis; Other autoimmune diseases such as Type I diabetes, autoimmune thyroid disorders, and Alzheimer's disease; Viral Diseases, such- as Epstein Barr Virus (EBV), Hepatitis B, Hepatitis C, HIV, HTLV 1, Varicella-Zoster Virus (VZV), Human Papilloma Virus (HPV); Cancer, such as fibrosarcoma, myκosarcoma, Iiposarcoma, chondrosarcoma, ostcogenic sarcoma, chordoma, angiosarcoma, endothcliosarcoma, lymphangiosaroo a, lymphangioendotheliosarcoma, synovioma, mesotheliorr-a, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cardnoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell cardnoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadcnocardnoma, medullary carcinoma, bronchogenic cardnoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, semϊnoma, embryonal cardnoma, Wilms' tumor, cervical cancer, testicular tumor, lung cardnoma, small cell lung carcinoma, bladder cardnoma, epithelial, cardnoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, and retinoblasto a, and cardnomas forming from tissue of the breast, prostate, kidney, bladder or colon, and ncoplastic disorders arising in adipose tissue, such as adipose cell tumors, e.g., lipomas, fibrolipomas, lipoblastomas, Upomatosis, hibemomas, hemangiomas and/or liposarcomas.

The use of at least one of the compounds of any one of dai s 1 to 4 in the preparation of a medicament for the treatment of a tyrosine kinase-assodated disease state.